Adsorption device and method for manufacturing the same

Abstract

To provide an adsorption device having improved separation performance by appropriately suppressing or preventing variation in particle size distribution within a column of hydroxyapatite powder packed in the column, and a method for manufacturing such an adsorption device. [Solution] The adsorption device of the present invention comprises a cylindrical column body having an adsorbent filling space therein, a first port provided at one end of the column body and having a first flow path through which liquid flows, a second port provided at the other end of the column body and having a second flow path through which liquid flows, and an adsorbent filled in the adsorbent filling space, wherein the adsorbent contains powder, and when the most frequent particle size in the particle size distribution of the powder present in a first region that is one end of the adsorbent filling space is A1 [μm] and the most frequent particle size in the particle size distribution of the powder present in a third region that is the other end of the adsorbent filling space is A3 [μm], the relationship 0.72≦A1 / A3≦1.38 is satisfied.

Description

[Technical field]

[0001] The present invention relates to an adsorption device having improved separation performance and a method for manufacturing such an adsorption device. [Background technology]

[0002] In recent years, hydroxyapatite has been widely used as a stationary phase material, i.e., an adsorbent, in chromatography for the purification and isolation of biopharmaceuticals such as antibodies and vaccines, for reasons such as its high biocompatibility and excellent safety.

[0003] An adsorption device having, as an adsorbent, a powder of hydroxyapatite (HAP) used as a stationary phase material in chromatography, is manufactured, for example, by applying the following manufacturing method.

[0004] That is, primary particles of hydroxyapatite are obtained by reacting a first liquid containing calcium hydroxide with a second liquid containing phosphoric acid while stirring, and a slurry containing the primary particles and their aggregates is dried and granulated to obtain secondary particles of hydroxyapatite.

[0005] The unsintered powder consisting of particles including these secondary particles is fired (sintered) to obtain a sintered powder.

[0006] Next, the unsintered or sintered hydroxyapatite powder is filled into the adsorbent filling space provided in the column body of the column, etc., to manufacture an adsorption device that includes the unsintered or sintered powder as a stationary phase material (adsorbent) (see, for example, Patent Document 1).

[0007] In the above-described method for manufacturing an adsorption device, the unsintered or sintered powder of hydroxyapatite serving as the adsorbent 3 is filled into the adsorbent filling space 20 provided in the column body 21 as described below.

[0008] That is, as shown in FIG. 7 , a column passing section 221 provided in an extension column 200 is connected to a column main body 21 provided in a column 2, and in a column combination 250 in which the column main body 21 and the column passing section 221 are connected, a slurry containing the adsorbent 3 is supplied from the upper end of the column passing section 221 to the adsorbent passing space 220 of the column passing section 221 with the column main body 21 facing vertically downward and the column passing section 221 facing vertically upward.

[0009] Then, while maintaining the column body 21 facing vertically downward and the column passing section 221 facing vertically upward, a filler liquid 66 such as water is pressurized and supplied from the upper end of the column passing section 221 using a pump 60, thereby causing the filler liquid 66 to pass from top to bottom through the extension column 200 and column 2.

[0010] Due to the passage of the filling liquid 66 from above to below in the extension column 200 and the column 2, and further due to gravity acting on the adsorbent 3, the adsorbent 3 contained in the slurry sequentially settles through the adsorbent passing space 220 of the column passing section 221 and the adsorbent filling space 20 of the column body 21, whereby the adsorbent 3 is filled in the adsorbent filling space 20 provided in the column body 21.

[0011] Here, as described above, the secondary particles of hydroxyapatite are obtained by granulating the primary particles and their aggregates, and therefore the hydroxyapatite powder (unsintered powder or sintered powder) used as the adsorbent 3 has a relatively large particle size distribution.

[0012] Therefore, when the adsorbent 3 is settled in the adsorbent passing space 220 of the column passing portion 221 and the adsorbent filling space 20 of the column body 21 and filled with the adsorbent 3, the adsorbent 3 having a large particle size is unevenly distributed on the lower side of the column 2, and the adsorbent 3 having a small particle size is unevenly distributed on the upper side of the column 2. That is, the particle size distribution of the adsorbent 3 in the column 2 varies. Therefore, due to the variation in the particle size distribution, problems occur such as the inability to impart column performance for purifying and isolating a biopharmaceutical as an adsorbent substance to an adsorption device equipped with the adsorbent 3, or the durability of the adsorbent 3 is reduced. The present inventor discovered such a problem. [Prior art documents] [Patent documents]

[0013] [Patent Document 1] JP 2011-68539 A Summary of the Invention [Problem to be solved by the invention]

[0014] It is an object of the present invention to provide an adsorption device having improved separation performance and a method for producing an adsorption device by which such an adsorption device can be produced. [Means for solving the problem]

[0015] Such an object can be achieved by the present invention described in the following (1) to (11). (1) a column body having a cylindrical shape and an adsorbent-filled space therein; a first port provided at one end of the column body and having a first flow path through which a liquid flows; a second port provided at the other end of the column body and having a second flow path through which a liquid flows; an adsorbent filled in the adsorbent filling space, The adsorbent includes a powder consisting of an aggregate of fine particles having a non-uniform particle size, An adsorption device characterized in that, when the most frequent particle size in the particle size distribution of the powder present in a first region that is one end of the adsorbent filling space is A1 [μm], and the most frequent particle size in the particle size distribution of the powder present in a third region that is the other end of the adsorbent filling space is A3 [μm], the relationship of 0.72≦A1 / A3≦1.38 is satisfied.

[0016] In this way, A1 / A3, which is a parameter expressing the uniformity of the most frequent particle sizes A1 and A3 of the adsorbent present in the first and third regions, which are both ends of the adsorbent-filled space, satisfies the relationship 0.72≦A1 / A3≦1.38. In other words, the lower limit of A1 / A3 is 0.72. The upper limit of A1 / A3 is 1.38. Therefore, it can be said that the adsorbent is filled in the adsorbent-filled space with a substantially uniform particle size distribution, and the column performance of purifying and isolating the adsorbed substance can be reliably imparted to the adsorption device equipped with this adsorbent. In addition, excellent durability can be imparted to the adsorbent equipped in the column.

[0017] (2) An adsorption device as described in (1) above, in which a sample liquid containing an adsorbent is supplied to the adsorbent filling space via the first flow path of the first lid part while the first lid part is positioned vertically above the second lid part, and the adsorption of the adsorbent is performed by the powder. The adsorption apparatus of the present invention is suitably applied to an adsorption apparatus having such a configuration.

[0018] (3) An adsorption device as described in (2) above, in which, when the most frequent particle size in the particle size distribution of the powder in a second region located at the midpoint between the one end and the other end is A2 [μm], 0.72≦A1 / A2≦1.38 is satisfied.

[0019] This means that the adsorbent is filled in the adsorbent filling space with a more uniform particle size distribution.

[0020] (4) The adsorption device according to (3) above, wherein when the frequency of the most frequent particle size in the particle size distribution of the powder in the second region is B2 (%), 0.80≦B1 / B2≦1.15 is satisfied.

[0021] This means that the adsorbent is filled in the adsorbent filling space with a more uniform particle size distribution.

[0022] (5) a first step of preparing a column including a cylindrical column body having an adsorbent-filled space therein, a first port provided at one end of the column body and having a first flow path through which a liquid flows, and a second port provided at the other end of the column body and having a second flow path through which a liquid flows; a second step of filling the adsorbent filling space of the column with a composition including an adsorbent containing a powder consisting of an aggregate of fine particles having a non-uniform particle size and a first liquid; and a third step of supplying a second liquid to the adsorbent filling space via the first flow path of the first port while the first port side of the column is positioned vertically downward.

[0023] According to the method for manufacturing an adsorption device of the present invention, the adsorbent filling space of the column body can be filled with the adsorbent with a substantially uniform particle size distribution.

[0024] (6) Prior to the second step, preparing an extension column including: a column passing section having a cylindrical shape and an adsorbent passing space therein; a cover body provided at one end of the column passing section and having a third flow path through which a liquid flows; and a connecting section provided at the other end of the column passing section and connectable to the one end side of the column main body; A method for manufacturing an adsorption device according to (5) above, comprising the step of connecting the other end of the column passing portion to the one end of the column body from which the first port is detached, via the connecting portion.

[0025] This allows the adsorbent to be packed in the adsorbent packing space of the column body with a substantially uniform particle size distribution.

[0026] (7) The adsorption apparatus obtained by the third step, The method for manufacturing an adsorption device according to (5) or (6) above, wherein, when the most frequent particle size in the particle size distribution of the powder present in one end region of the adsorbent filling space is A1 [μm] and the most frequent particle size in the particle size distribution of the powder present in the other end region of the adsorbent filling space is A3 [μm], 0.72≦A1 / A3≦1.38 is satisfied.

[0027] According to the manufacturing method of the adsorption device of the present invention, the adsorbent can be filled into the adsorbent filling space of the column body with an almost uniform particle size distribution, and specifically, the mode particle sizes A1 and A3 can satisfy the relationship 0.72≦A1 / A3≦1.38. In other words, the lower limit of A1 / A3 is 0.72, and the upper limit of A1 / A3 is 1.38. Effect of the Invention

[0028] The adsorption device of the present invention has improved separation performance, and the manufacturing method of the present invention can manufacture such an adsorption device. [Brief description of the drawings]

[0029] [Figure 1] 1 is a vertical cross-sectional view showing an example of an adsorption device according to an embodiment of the present invention. [Diagram 2] 4 is a flowchart illustrating a method for manufacturing an adsorption device according to an embodiment of the present invention. [Diagram 3] 5A to 5C are vertical cross-sectional views for explaining a method for manufacturing a suction device according to an embodiment of the present invention. [Figure 4] 5A to 5C are vertical cross-sectional views for explaining a method for manufacturing a suction device according to an embodiment of the present invention. [Diagram 5] 5A to 5C are vertical cross-sectional views for explaining a method for manufacturing a suction device according to an embodiment of the present invention. [Figure 6] 4 is a graph showing the particle size distribution of the adsorbent present at a position corresponding to a first region in the adsorption device of Example 1. [Figure 7] 11A and 11B are vertical cross-sectional views for explaining a conventional method for manufacturing a suction device. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An adsorption apparatus and a method for manufacturing an adsorption apparatus according to the present invention will now be described in detail with reference to preferred embodiments shown in the accompanying drawings.

[0031] <Adsorption device> Fig. 1 is a vertical cross-sectional view showing an example of an adsorption device according to an embodiment of the present invention. In the following description, the upper side in Fig. 1 is referred to as the "inlet side" and the lower side is referred to as the "outlet side".

[0032] Here, the "inlet side" refers to the side where liquids such as a sample liquid (liquid containing the adsorbent) and an eluent (phosphate buffer solution, water, etc.) are supplied into the adsorption device when separating (purifying) the adsorbent as an isolated product such as a biopharmaceutical or the like to be separated, i.e., a protein, etc., while the "outlet side" refers to the side opposite the inlet side, i.e., the side from which the liquid flows out of the adsorption device.

[0033] An adsorption device 1 shown in FIG. 1, which separates (purifies) isolated substances such as proteins from a sample liquid, comprises a column 2, a granular adsorbent (packing material) 3, and two filter members 4 and 5.

[0034] The column 2 has a column body 21, and a lid part 22 (first port) and a lid part 23 (second port) attached to the inlet end and the outlet end of the column body 21, respectively.

[0035] The column body 21 is composed of, for example, a cylindrical member. Examples of materials for the components (members) constituting the column 2 including the column body 21 include various glass materials, various resin materials, various metal materials, various ceramic materials, and the like.

[0036] The column body 21 has filter members 4, 5 disposed so as to close the inlet and outlet openings thereof, respectively, and has lids 22, 23 attached to the inlet and outlet ends thereof, respectively.

[0037] In the column 2 thus configured, an adsorbent filling space 20 is defined by the column body 21 and the filter members 4 and 5. Furthermore, the column body 21 and the lids 22 and 23 ensure liquid-tightness in the adsorbent filling space 20. At least a portion of the adsorbent filling space 20 (in this embodiment, almost to the full extent) is filled with the adsorbent 3.

[0038] The inner diameter (column inner diameter) of the adsorbent filling space 20 is appropriately set according to the volume of the sample liquid, and is not particularly limited, but is preferably about 3.0 mm to 70.0 mm, and more preferably about 5.0 mm to 50.0 mm. The length (column length) of the adsorbent filling space 20 is, for example, preferably about 10.0 mm to 300.0 mm, and more preferably about 20.0 mm to 200.0 mm. By using a column 2 having such dimensions (inner diameter and length) of the adsorbent filling space 20 for isolating an isolated substance contained in a sample liquid, the isolated substance can be purified with excellent accuracy.

[0039] By setting the dimensions of the adsorbent filling space 20 as described above and by setting the dimensions of the adsorbent 3 described below as well, it is possible to selectively isolate (purify) the desired isolate from the sample liquid, i.e., to reliably separate the isolate (adsorbed substance) such as protein from impurities other than the isolate contained in the sample liquid.

[0040] The isolated substances isolated (purified) using the adsorbent 3 are not limited to proteins such as acidic proteins, such as albumin and antibodies, and basic proteins, but include, for example, negatively charged substances such as acidic amino acids, DNA, RNA, and negatively charged liposomes, as well as positively charged substances such as basic amino acids, positively charged cholesterol, and positively charged liposomes. In other words, various substances such as antibodies and biopharmaceuticals such as vaccines can be purified and isolated using the adsorbent 3 as adsorbents.

[0041] Moreover, by attaching each of the lids 22, 23 to the column body 21, liquid-tightness between them is ensured.

[0042] The lid portion 22 (first port) and the lid portion 23 (second port) each have a cap 28 and a cap 29, an inlet pipe 24 (first flow path) and an outlet pipe 25 (second flow path), and a lid 26 and a lid 27, respectively.

[0043] The caps 28, 29 are attached by screwing to the inlet end (one end) and the outlet end (the other end) of the column body 21, respectively, with the inlet end of the column body 21 vertically upward and the outlet end vertically downward, and the caps 28, 29 and the column body 21 ensure liquid-tightness between them.

[0044] The inlet pipe 24 and the outlet pipe 25 are constituted by a pipe body through which the liquid flows, and are fixed (fixed) in a liquid-tight manner to the approximate center of each of the caps 28 and 29. The lids 26 and 27 are disposed between the filter members 4 and 5 and the caps 28 and 29, and have flow paths 41 and 51 communicating with the inlet pipe 24 and the outlet pipe 25, respectively. The liquid is supplied to the adsorbent 3 through the inlet pipe 24, the lid 26, and the filter member 4. The sample liquid supplied to the adsorbent 3 passes between the adsorbents 3 (gaps) and flows out of the column 2 through the filter member 5, the lid 27, and the outlet pipe 25. At this time, the isolated substance (adsorbed substance) contained in the sample liquid (sample) and impurities other than the isolated substance are separated based on the difference in adsorptivity to the adsorbent 3 and the difference in affinity to the eluent.

[0045] That is, the adsorption device 1 supplies the liquid to the adsorbent filling space 20 via the inlet pipe 24 with the lid portion 22 positioned vertically above the lid portion 23, adsorbs the isolated matter using the adsorbent 3 filled in the adsorbent filling space 20, and uses this adsorption of the isolated matter to separate the isolated matter.

[0046] Each of the filter members 4, 5 has a function of preventing the adsorbent 3 from flowing out of the adsorbent filling space 20, that is, a function of retaining the adsorbent 3 within the adsorbent filling space 20. These filter members 4, 5 are each composed of, for example, a polypropylene mesh, a sintered polyethylene particle filter, a stainless steel mesh filter, or a sintered stainless steel particle filter.

[0047] In the adsorption device 1, the adsorbent 3 includes a powder consisting of an aggregate of fine particles having a non-uniform particle size, and has an ability to adsorb isolated substances (adsorbed substances) contained in a sample liquid (sample). For example, it is preferable that the adsorbent 3 is composed of unsintered powder consisting of particles including secondary particles of hydroxyapatite, or sintered powder thereof. In this specification, the unsintered powder and sintered powder of hydroxyapatite are collectively referred to as "hydroxyapatite powder". Therefore, in the following, a case where hydroxyapatite powder is filled in the adsorbent filling space 20 of the column body 21 as the adsorbent 3 will be described as an example.

[0048] In the present invention, in the adsorbent filling space 20 provided in the adsorption device 1, when the most frequent particle size in the particle size distribution of the adsorbent 3 present in the first region a1 is A1 [μm] and the most frequent particle size in the particle size distribution of the adsorbent 3 present in the third region a3 is A3 [μm], the adsorbent 3 satisfies the relationship 0.72≦A1 / A3≦1.38. However, since an adsorption device 1 satisfying this relationship can be obtained by applying a manufacturing method for an adsorption device described later, this relationship will be explained later.

[0049] In the present embodiment, the adsorbent 3 filled in the adsorbent filling space 20 of the adsorption device 1 according to the embodiment of the present invention is a powder of hydroxyapatite, but this adsorbent 3 includes a powder consisting of an aggregate of fine particles having non-uniform particle diameters. In a preferred embodiment of the present invention, the aggregate of fine particles preferably has a predetermined most frequent particle diameter (mode diameter) and has a mountain-shaped particle diameter distribution centered on the value of the most frequent particle diameter. In another preferred embodiment of the present invention, the aggregate of fine particles preferably has particle diameters distributed above and below the value of the most frequent particle diameter such that the frequency of occurrence decreases the further away from the value of the most frequent particle diameter.

[0050] The powder constituting the adsorbent 3 filled in the adsorbent filling space 20 of the adsorption device 1 according to the embodiment of the present invention may be, in addition to the hydroxyapatite powder shown in this embodiment, a powder of an inorganic compound such as silica, fluoroapatite, calcium pyrophosphate, magnesium pyrophosphate, alumina, zirconia, etc., a powder of an organic compound such as polymer beads, a polymer such as sepharose or agarose, or a mixture of these. The effect of the present invention is more effective when the adsorbent 3 contains a powder consisting of an aggregate of fine particles having non-uniform particle diameters.

[0051] The hydroxyapatite powder may be a commercially available product, such as CHT (USA registered trademark) Type I, CHT (USA registered trademark) Type II, or CHT (USA registered trademark) XT, MPC, or CFT, manufactured by Bio-Rad.

[0052] When producing hydroxyapatite powder, the hydroxyapatite powder can be produced according to the method described in, for example, JP-B-7-88205 and Japanese Patent No. 5724050. Specifically, in order to produce hydroxyapatite powder, a production method can be carried out that includes a step [S1A] of reacting a first liquid containing a calcium source such as calcium hydroxide with a second liquid containing a phosphorus source such as phosphoric acid while stirring to obtain a slurry containing primary particles of hydroxyapatite and their aggregates, a step [S2A] of physically crushing the aggregates contained in the slurry and dispersing the crushed aggregates in the slurry, and a step [S3A] of drying the slurry and granulating the crushed aggregates to obtain a powder mainly composed of particles containing secondary particles of hydroxyapatite.

[0053] The powder constituting the adsorbent 3 filled in the adsorbent filling space 20 of the adsorption device 1 according to the embodiment of the present invention may be an unsintered powder or a sintered powder. The powder constituting the adsorbent 3 filled in the adsorbent filling space 20 of the adsorption device 1 according to the embodiment of the present invention may be classified or unclassified. When the powder is unclassified, the powder has a wider particle size distribution, and therefore the effect of the present invention may be more significantly exhibited.

[0054] The powder constituting the adsorbent 3 filled in the adsorbent filling space 20 of the adsorption device 1 according to an embodiment of the present invention, when classified, may have an average particle size and particle size distribution of 20±4 μm, 40±4 μm, 60±4 μm, 80±4 μm, etc.

[0055] The above powder is filled as the adsorbent 3 (stationary phase material) into the adsorbent filling space 20 of the column body 21 to obtain the adsorption device 1. When filling the adsorbent filling space 20 with the adsorbent 3, a manufacturing method for an adsorption device according to an embodiment of the present invention is applied.

[0056] <Method of manufacturing the adsorption device> Hereinafter, a method for manufacturing the suction device 1 to which the manufacturing method for the suction device according to the embodiment of the present invention is applied will be described.

[0057] Fig. 2 is a flow chart showing a method for manufacturing an adsorption device according to an embodiment of the present invention, and Figs. 3 to 5 are vertical cross-sectional views for explaining the method for manufacturing an adsorption device according to an embodiment of the present invention. In the following description, the upper side in Figs. 3 to 5 is called the "inlet side" or "upper side", and the lower side is called the "outlet side" or "lower side".

[0058] As shown in FIG. 2, a manufacturing method of an adsorption device according to an embodiment of the present invention includes a step [S1B] of preparing a column 2, a step [S2B] of preparing an extension column 200 and connecting the column passage section 221 of the extension column 200 to the column body 21 of the column 2 to obtain a column combination body 250, a step [S3B] of filling the column combination body 250 with a slurry 30 containing hydroxyapatite powder and a dispersion liquid that constitutes the adsorbent 3, a step [S4B] of supplying a filling liquid 66 from the extension column 200 side to the column body 21 side in the column combination body 250 while the column combination body 250 is oriented so that the extension column 200 is vertically downward, and a step [S5B] of detaching the extension column 200 from the column combination body 250 to obtain an adsorption device 1.

[0059] These steps will be described in order below. [S1B: Preparation of column 2] In this step, the column 2 included in the above-mentioned adsorption apparatus 1 is prepared (first step).

[0060] In this embodiment, as shown in Figure 3, the column 2 is prepared in such a state that, of the lid portions 22 and 23 (ports), the lid portion 23 is attached to the outlet end of the column body 21 and the lid portion 22 is not attached to the inlet end of the column body 21.

[0061] [S2B: Step of preparing column connector 250] In this step, an extension column 200 is prepared, and the column passage portion 221 of the extension column 200 is connected to the column body 21 of the column 2 to obtain a column combination 250 (see FIG. 4).

[0062] As shown in FIG. 4, the extension column 200 has a column passing portion 221, and a cap 228 and a connecting portion 229 attached to the inlet end (one end) and outlet end (the other end) of the column passing portion 221, respectively.

[0063] The column passing section 221 is cylindrical, has an adsorbent passing space 220 therein, and has substantially the same configuration as the column body 21 that the column 2 comprises.

[0064] A cap 228 having an inlet pipe 224 (third flow path) at its center can be attached to the inlet end of the column passing section 221 by screwing, but is left unattached in this step [S2B]. In addition, a connecting section 229 connectable to the inlet end of the column body 21 is attached to the outlet end of the column passing section 221 by screwing.

[0065] Then, by screwing this connecting part 229 onto the inlet end of the column body 21 of the column 2, a column connection body 250 can be obtained in which the outlet end of the column passing part 221 provided in the extension column 200 is connected to the inlet end of the column body 21 provided in the column 2 via the connecting part 229.

[0066] [S3B: Step of supplying slurry 30 to column connection body 250] In this step, the column connection body 250 is filled with a slurry 30 containing hydroxyapatite powder as a powder consisting of an aggregate of fine particles having non-uniform particle diameters that constitute the adsorbent 3, and a dispersion liquid (first liquid) (second step).

[0067] This process includes a step [S3B-1] of preparing a slurry 30 containing hydroxyapatite powder constituting the adsorbent 3 and a dispersion liquid (first liquid), and a step [S3B-2] of supplying the slurry 30 to the column combination 250.

[0068] Hereinafter, steps [S3B-1] and [S3B-2] constituting this step [S3B] will be described in order.

[0069] [S3B-1: Step of preparing slurry 30 containing adsorbent 3 and dispersion liquid] In this process, the powder (unsintered powder or sintered powder) obtained by the manufacturing method for hydroxyapatite powder is stirred as an adsorbent 3 in a dispersion liquid (first liquid) to prepare a slurry 30 (composition) in which the adsorbent 3 is dispersed in the dispersion liquid.

[0070] Specifically, for example, while the dispersion liquid is stirred in a container (not shown), hydroxyapatite powder is mixed into the dispersion liquid, and by this mixing, the hydroxyapatite powder (adsorbent 3) is dispersed in the dispersion liquid.

[0071] The dispersion liquid (first liquid) in which the hydroxyapatite powder is dispersed is water or water containing a buffer solution or salts, and examples of the buffer solution or salts include sodium salts, potassium salts, ammonium salts, magnesium salts, calcium salts, chlorides, fluorides, acetates, phosphates, and citrates, and one or more of these can be used in combination. Among these, sodium phosphate buffer is preferable. By using a sodium phosphate buffer as the dispersion liquid, the hydroxyapatite powder can be stably dispersed in the slurry.

[0072] Furthermore, when a sodium phosphate buffer is used, its concentration is not particularly limited, but is preferably set to about 0.1 M or more and 0.6 M or less, more preferably about 0.15 M or more and 0.4 M or less.

[0073] Furthermore, the pH of the sodium phosphate buffer is preferably about 6.0 or more and about 9.0 or less, and more preferably about 6.8 or more and about 9.0 or less.

[0074] The liquid temperature of the dispersion liquid is not particularly limited, but is preferably set to about 0°C or higher and 70°C or lower, and more preferably about 25°C or higher and 50°C or lower.

[0075] [S3B-2: Step of supplying slurry 30 to column connection body 250] In this step, the slurry 30 prepared in the above step [S3B-1] is supplied to the column combination 250.

[0076] Specifically, as shown in FIG. 4, in a column connection body 250 in which the column body 21 of the column 2 and the column passing portion 221 of the extension column 200 are connected, the column body 21 of the column 2 is positioned vertically downward, and the column passing portion 221 of the extension column 200 is positioned vertically upward.

[0077] Then, the slurry 30 in which the adsorbent 3 is dispersed is supplied to the adsorbent passing space 220 provided in the column passing section 221 of the extension column 200 from an opening that opens at the inlet side end (upper end) of the column passing section 221.

[0078] This allows the slurry 30 containing the adsorbent 3 to be filled in the adsorbent filling space 20 provided in the column body 21 of the column 2 to be filled in the adsorbent passing space 220 provided in the column passing section 221 of the column combination 250 to which the column body 21 is connected. Note that the adsorbent 3 filled in the adsorbent passing space 220 of the column passing section 221 may move down within the adsorbent passing space 220 due to its own weight, and a part of it may move to the adsorbent filling space 20 of the column body 21.

[0079] [S4B: Step of supplying the packing liquid 66 to the column connection body 250] In this step, the column combination 250 is oriented such that the extension column 200 faces vertically downward, and the filling liquid 66 (second liquid) is supplied to the column combination 250 from the extension column 200 side (third step).

[0080] This process includes a step [S4B-1] of positioning the column connector 250 so that the extension column 200 is vertically downward, and a step [S4B-2] of supplying the filler liquid 66 (second liquid) from the extension column 200 side to the column 2 (column body 21) side of the column connector 250.

[0081] Hereinafter, steps [S4B-1] and [S4B-2] constituting this step [S4B] will be described in order.

[0082] [S4B-1: Step of arranging the column connector 250 with the extension column 200 side vertically downward] In this step, the column combination 250 in which the adsorbent passing space 220 of the column passing section 221 of the extension column 200 is filled with the slurry 30 is oriented so that the extension column 200 side faces vertically downward.

[0083] Specifically, in a column combination 250 in which the adsorbent passing space 220 of the column passing section 221 of the extension column 200 is filled with slurry 30, a cap 228 is attached to the inlet end (upper end) of the column passing section 221 of the extension column 200 by screwing.

[0084] 5, the column coupler 250 in which the column body 21 of the column 2 and the column passing part 221 of the extension column 200 are connected is disposed in a position in which the column body 21 of the column 2 is vertically upward and the column passing part 221 of the extension column 200 is vertically downward. That is, in the column coupler 250 in which the column 2 and the extension column 200 are connected, the column coupler 250 is disposed in a position in which the inlet side end of the column coupler 250 is vertically downward and the outlet side end of the column coupler 250 is vertically upward.

[0085] When arranging the column coupler 250 in this manner, the filling liquid 66 (second liquid) does not necessarily have to be supplied to the adsorbent passing space 220 of the column passing section 221 via the inflow pipe 224 of the cap 228, but it is preferable to supply the filling liquid 66 to the adsorbent passing space 220 in advance. This makes it possible to appropriately suppress or prevent the adsorbent 3 filled in the adsorbent passing space 220 from unintentionally flowing down to the inflow end side of the column passing section 221, i.e., the cap 228 side, when the column coupler 250 is inverted so that the extension column 200 is vertically downward from the vertically upward position, as shown in Figures 4 and 5 .

[0086] When the filling liquid 66 is supplied to the adsorbent passing space 220 in advance, the column coupler 250 is oriented such that the column body 21 of the column 2 is vertically downward and the column passing section 221 of the extension column 200 is vertically upward, and the supply of the filling liquid 66 is started to the adsorbent passing space 220 of the column passing section 221 through the inflow pipe 224 of the cap 228. After that, when the adsorbent passing space 220 of the column passing section 221 and the adsorbent filling space 20 of the column body 21 are almost fully supplied with the filling liquid 66, it is preferable to invert the column coupler 250 so that the column body 21 of the column 2 is vertically upward and the column passing section 221 of the extension column 200 is vertically downward. This makes it possible to more accurately suppress or prevent the adsorbent 3 filled in the adsorbent passing space 220 from unintentionally flowing down to the inflow end side of the column passing section 221.

[0087] [S4B-2: Step of supplying the packing liquid 66 to the column connection body 250 from the extension column 200 side] In this step, the packing liquid 66 (second liquid) is supplied from the column passing portion 221 side of the extension column 200 to the column body 21 side of the column 2 in the column connection body 250. That is, the packing liquid 66 (second liquid) is supplied from the inlet end side to the outlet end side of the column connection body 250.

[0088] Specifically, for example, as shown in FIG. 5, the column connection body 250 is oriented such that the column body 21 of the column 2 is vertically upward and the column passing section 221 of the extension column 200 is vertically downward. Then, using a pump 60, the filling liquid 66 stored in a container 65 is supplied to the adsorbent passing space 220 of the column passing section 221 via the inlet pipe 224 (third flow path) of the cap 228.

[0089] By supplying the filling liquid 66 to the adsorbent passing space 220 of the column passing section 221, the filling liquid 66 fills the adsorbent passing space 220 of the column passing section 221 and the adsorbent filling space 20 of the column body 21 until they are almost full, and then is discharged from the outflow pipe 25 of the lid section 23. That is, by the driving force of the pump 60, the filling liquid 66 flows and passes through the column connected body 250, with the adsorbent passing space 220 of the column passing section 221 on the upstream side and the adsorbent filling space 20 of the column body 21 on the downstream side.

[0090] Within the adsorbent passing space 220 of the column passing section 221 and the adsorbent filling space 20 of the column body 21, the filling liquid 66 passes through the adsorbent passing space 220 upstream and the adsorbent filling space 20 downstream, i.e., the filling liquid 66 passes from the bottom to the top of the column connection body 250, whereby the adsorbent 3 contained in the slurry rises sequentially through the adsorbent passing space 220 of the column passing section 221 and the adsorbent filling space 20 of the column body 21, and the adsorbent 3 is filled in the adsorbent filling space 20 of the column body 21.

[0091] When the adsorbent 3 rises in the adsorbent passing space 220 and the adsorbent filling space 20, the adsorbent 3 is subjected to a downward force due to gravity in addition to an upward force acting on the adsorbent 3 due to the passing filling liquid 66.

[0092] Here, the adsorbent 3, i.e., hydroxyapatite powder, contains primary particles and secondary particles obtained by granulating the aggregates thereof, and therefore has a relatively wide particle size distribution. Therefore, it can be said that the hydroxyapatite powder is a powder consisting of an aggregate of fine particles having non-uniform particle sizes.

[0093] In this way, even if the adsorbent 3 has a relatively large particle size distribution, when the adsorbent 3 is filled in the adsorbent filling space 20 provided in the column body 21, both an upward force due to the filling liquid 66 passing through the adsorbent passing space 220 and the adsorbent filling space 20 and a downward force due to gravity act on the adsorbent 3. Therefore, the adsorbent 3 rises in the adsorbent passing space 220 and the adsorbent filling space 20 while convecting in the adsorbent passing space 220 and the adsorbent filling space 20, and is filled in the adsorbent filling space 20. Therefore, the adsorbent 3 is filled in the adsorbent filling space 20 while suppressing or preventing the adsorbent 3 having a large particle size from being unevenly distributed on the lower side of the adsorbent filling space 20 and the adsorbent 3 having a small particle size from being unevenly distributed on the upper side of the adsorbent filling space 20. Therefore, the adsorbent 3 is filled in the adsorbent filling space 20 while appropriately suppressing or preventing the occurrence of variations in the particle size distribution of the adsorbent 3 in the adsorbent filling space 20 (column 2). Therefore, since the variation in particle size distribution is appropriately suppressed or prevented, the column performance for purifying and isolating the biopharmaceutical, i.e., the adsorption substance, can be reliably imparted to the adsorption device 1 equipped with the adsorbent 3. In addition, excellent durability can be imparted to the adsorbent 3 equipped in the column 2.

[0094] By supplying the filling liquid 66 to the column connection body 250 in this manner, the adsorbent 3 is filled into the adsorbent filling space 20 with variation in its particle size distribution appropriately suppressed or prevented, that is, the adsorbent filling space 20 is filled with a substantially uniform particle size distribution. In the present invention, the degree of the particle size distribution satisfies the relationship of 0.72≦A1 / A3≦1.38, specifically, as shown in FIG. 1, when the region at the inflow side end (one end) of the adsorbent filling space 20 is defined as a first region a1, the region at the center (midpoint) between the inflow side end and the outflow side end is defined as a second region a2, and the region at the outflow side end (other end) is defined as a third region a3, where A1 [μm] is the most frequent particle size in the particle size distribution of the adsorbent 3 present in the first region a1 and A3 [μm] is the most frequent particle size in the particle size distribution of the adsorbent 3 present in the third region a3. In this way, A1 / A3, which is a parameter expressing the uniformity of the most frequent particle diameters A1 and A3 of the adsorbent 3 present in the first region a1 and the third region a3, which are both ends of the adsorbent filling space 20, satisfies the relationship of 0.72≦A1 / A3≦1.38. Therefore, it can be said that the adsorbent 3 is filled in the adsorbent filling space 20 with a substantially uniform particle size distribution, and the adsorption device 1 including the adsorbent 3 can be reliably provided with column performance for purifying and isolating the biopharmaceutical, i.e., the adsorbent substance. In other words, the adsorption device 1 including the adsorbent 3 can be made to have improved separation performance. In addition, excellent durability can be provided to the adsorbent 3 included in the column 2. Furthermore, the A1 / A3 preferably satisfies the relationship of 0.85≦A1 / A3≦1.20. It is more preferable that the A1 / A3 satisfies the relationship of 0.90≦A1 / A3≦1.10. This makes it possible to more significantly exhibit the above-mentioned effect. In other words, the lower limit of the A1 / A3 ratio is 0.72, more preferably 0.85, and even more preferably 0.90, and the upper limit of the A1 / A3 ratio is 1.38, more preferably 1.20, and even more preferably 1.10.

[0095] In addition, the relationship between the most frequent particle diameters A1 and A3 of the adsorbent 3 present in the first region a1 and the third region a3 is preferably such that |A1-A3|, which is another parameter expressing the uniformity of the adsorbent 3 present in the first region a1 and the third region a3, satisfies the relationship |A1-A3|≦10 μm, and more preferably satisfies the relationship 0.1 μm≦|A1-A3|≦4.0 μm, in addition to the A1 / A3 relationship. By satisfying such a relationship, it can be said that the adsorbent 3 is filled in the adsorbent filling space 20 with a substantially uniform particle size distribution, and the adsorption device 1 equipped with the adsorbent 3 can be reliably provided with the column performance for purifying and isolating the biopharmaceutical, i.e., the adsorbent substance. In addition, excellent durability can be provided to the adsorbent 3 equipped in the column 2. In other words, the lower limit value of |A1-A3| is more preferably 0.1 μm. Moreover, the upper limit of |A1-A3| is preferably 10 μm, and more preferably 4.0 μm.

[0096] Furthermore, when the frequency of the most frequent particle size A1 in the particle size distribution of the adsorbent 3 present in the first region a1 is B1 (%), and the frequency of the most frequent particle size A3 in the particle size distribution of the adsorbent 3 present in the third region a3 is B3 (%), it is preferable that 0.9≦B1 / B3≦1.1 is satisfied, and it is more preferable that 0.95≦B1 / B3≦1.05 is satisfied. In other words, the lower limit value of B1 / B3 is preferably 0.9, and more preferably 0.95. Moreover, the upper limit value of B1 / B3 is preferably 1.1, and more preferably 1.05.

[0097] Moreover, the relationship between the frequencies B1 and B3 of the most frequent particle sizes A1 and A3 of the adsorbent 3 present in the first region a1 and the third region a3, other than the B1 / B3 relationship, is preferably such that |B1-B3|, which is another parameter expressing the uniformity of the adsorbent 3 present in the first region a1 and the third region a3, satisfies the relationship |B1-B3|≦3.0%, and more preferably satisfies the relationship 0.5%≦|B1-B3|≦3.0%. In other words, the lower limit of |B1-B3| is more preferably 0.5%. Moreover, the upper limit of |B1-B3| is preferably 3.0%.

[0098] When the frequencies B1, B3 of the most frequent particle sizes A1, A3 of the adsorbent 3 present in the first region a1 and the third region a3 satisfy the above-mentioned relationship, it can be said that the adsorbent 3 is filled in the adsorbent filling space 20 with a more uniform particle size distribution, and the above-mentioned effect can be exhibited more significantly.

[0099] Furthermore, in addition to the relationship between the most frequent particle diameters A1 and A3 of the adsorbent 3 present in the first region a1 and the third region a3, when the most frequent particle diameter in the particle size distribution of the adsorbent 3 present in the second region a2 is A2 [μm], the relationship between the most frequent particle diameters A1 and A2 preferably satisfies the relationship of 0.72≦A1 / A2≦1.38. The A1 / A2 is more preferably 0.85≦A1 / A2≦1.2, more preferably 0.90≦A1 / A2≦1.1, and most preferably 0.95≦A1 / A2≦1.05. In other words, the lower limit of A1 / A2 is preferably 0.72, more preferably 0.85, more preferably 0.90, and most preferably 0.95. The upper limit of A1 / A2 is preferably 1.38, more preferably 1.2, more preferably 1.1, and most preferably 1.05.

[0100] Moreover, the relationship between the most frequent particle sizes A1 and A2 of the adsorbent 3 present in the first region a1 and the second region a2 is preferably such that, in addition to the A1 / A2 relationship, |A1-A2|, which is another parameter expressing the uniformity of the adsorbent 3 present in the first region a1 and the second region a2, satisfies the relationship |A1-A2|≦10 μm, and more preferably satisfies the relationship 0.1 μm≦|A1-A2|≦4.0 μm. In other words, the lower limit of |A1-A2| is more preferably 0.1 μm. The upper limit of |A1-A2| is preferably 10 μm, and more preferably 4.0 μm.

[0101] When the most frequent particle sizes A1, A2 of the adsorbent 3 present in the first region a1 and the second region a2 satisfy the above-mentioned relationship, it can be said that the adsorbent 3 is filled in the adsorbent filling space 20 with a more uniform particle size distribution, and the above-mentioned effect can be exhibited more significantly.

[0102] Furthermore, in addition to the relationship between the frequencies B1 and B3 of the most frequent particle diameters A1 and A3 of the adsorbent 3 present in the first region a1 and the third region a3, when the frequency of the most frequent particle diameter A2 in the particle size distribution of the adsorbent 3 present in the second region a2 is B2 (%), B1 / B2 is preferably 0.80≦B1 / B2≦1.15, more preferably 0.85≦B1 / B2≦1.10, even more preferably 0.90≦B1 / B2≦1.05, and even more preferably 0.95≦B1 / B2≦1.02. In other words, the lower limit of B1 / B2 is preferably 0.80, more preferably 0.85, even more preferably 0.90, and even more preferably 0.95. The upper limit of B1 / B2 is preferably 1.15, more preferably 1.10, even more preferably 1.05, and even more preferably 1.02.

[0103] Moreover, the relationship between the frequencies B1 and B2 of the most frequent particle sizes A1 and A2 of the adsorbent 3 present in the first region a1 and the second region a2, other than the B1 / B2 relationship, is preferably such that |B1-B2|, which is another parameter expressing the uniformity of the adsorbent 3 present in the first region a1 and the second region a2, satisfies the relationship |B1-B2|≦3.0%, and more preferably satisfies the relationship 0.5%≦|B1-B2|≦3.0%. In other words, the lower limit of |B1-B3| is more preferably 0.5%. Moreover, the upper limit of |B1-B3| is preferably 3.0%.

[0104] When the frequencies B1, B2 of the most frequent particle sizes A1, A2 of the adsorbent 3 present in the first region a1 and the second region a2 satisfy the above-mentioned relationship, it can be said that the adsorbent 3 is filled in the adsorbent filling space 20 with a more uniform particle size distribution, and the above-mentioned effect can be exhibited more significantly.

[0105] When the median particle diameters of the adsorbent 3 present in the first region a1, the second region a2 and the third region a3 are C1, C2 and C3, respectively, it is preferable that C1 / C3 satisfies the formula 0.7≦C1 / C3≦1.3, it is more preferable that C1 / C3 satisfies the formula 0.8≦C1 / C3≦1.2, and it is even more preferable that C1 / C3 satisfies the formula 0.9≦C1 / C3≦1.1.

[0106] Furthermore, when the median particle sizes of the adsorbent 3 present in the first region a1, the second region a2 and the third region a3 are C1, C2 and C3, respectively, it is preferable that C1, C2 and C3 satisfy 0.9≦C1 / C3≦1.1 and 0.9≦C1 / C2≦1.1, and it is more preferable that C1, C2 and C3 satisfy 0.95≦C1 / C2≦1.05 and 0.95≦C1 / C2≦1.05.

[0107] Furthermore, when the average particle sizes of the adsorbent 3 present in the first region a1, the second region a2, and the third region a3 are D1, D2, and D3, respectively, it is preferable that D1, D2, and D3 satisfy 0.9≦D1 / D3≦1.1 and 0.9≦D1 / D2≦1.1, and it is more preferable that D1, D2, and D3 satisfy 0.95≦D1 / D3≦1.05 and 0.95≦D1 / D2≦1.05.

[0108] Furthermore, when the standard deviations of the particle sizes of the adsorbent 3 present in the first region a1, the second region a2 and the third region a3 are E1, E2 and E3, respectively, it is preferable that E1, E2 and E3 satisfy 0.9≦E1 / E3≦1.1 and 0.9≦E1 / E2≦1.1, and it is more preferable that E1, E2 and E3 satisfy 0.95≦E1 / E3≦1.05 and 0.95≦E1 / E2≦1.05.

[0109] Furthermore, when the particle size ranges of the adsorbent 3 present in the first region a1, the second region a2 and the third region a3 are F1, F2 and F3, respectively, it is preferable that they satisfy 0.9≦F1 / F3≦1.1 and 0.9≦F1 / F2≦1.1, and it is more preferable that they satisfy 0.95≦F1 / F3≦1.05 and 0.95≦F1 / F2≦1.05.

[0110] As described above, it can be said that by making not only the mode particle size of the adsorbent 3 present in the first region a1, the second region a2, and the third region a3, but also the median particle size, average particle size, standard deviation of the particle sizes, and particle size range of the adsorbent 3 satisfy the above-mentioned relationships, the occurrence of variation in the particle size distribution of the adsorbent 3 filled in the adsorbent filling space 20 (column 2) is appropriately suppressed or prevented. Therefore, by satisfying the above-mentioned relationships, the above-mentioned effect can be more significantly exhibited.

[0111] The filling liquid 66 (second liquid) supplied to the column combination 250 may be the same as the dispersion liquid contained in the slurry 30 described above, and among them, water is preferable. Water is easier to handle than a buffer solution or the like, and is therefore preferably used as the filling liquid 66 supplied in large quantities to the column combination 250 using the pump 60.

[0112] The temperature of the filling liquid 66 is not particularly limited, but is preferably set to about 0°C or higher and 70°C or lower, and more preferably about 25°C or higher and 50°C or lower.

[0113] Furthermore, the flow rate when the filling liquid 66 is supplied to the column connection body 250 is not particularly limited, but is preferably set to about 70 mL / min or more, more preferably about 100 mL / min to 250 mL / min, and even more preferably about 110 mL / min to 180 mL / min.

[0114] The time for supplying the packing solution 66 to the column connection assembly 250 is not particularly limited, but is preferably set to about 5 minutes or more, and more preferably from 20 minutes to 60 minutes.

[0115] By setting the conditions for supplying the packing liquid 66 to the column connection body 250 as described above, the adsorbent 3 can be packed into the adsorbent filling space 20 of the column body 21, the dimensions of which are set as described above, while accurately suppressing or preventing variations in the particle size distribution. That is, the adsorbent 3 can be packed into the adsorbent filling space 20 while reliably satisfying the relationship 0.72≦A1 / A3≦1.38, where A1 / A3 is a parameter that represents the uniformity of the most frequent particle sizes A1 and A3 of the adsorbent 3.

[0116] [S5B: Step of removing the extension column 200] In this step, the extension column 200 is detached from the column connector 250 to obtain the adsorption apparatus 1.

[0117] Specifically, the connecting part 229 that is attached by screwing to the inlet end of the column body 21 of the column 2 is detached from the column body 21. As a result, the extension column 200 is detached from the column 2 in the column connector 250.

[0118] Then, the lid part 22 is attached to the inlet end part of the column body 21 of the column 2 from which the connecting part 229 has been detached, by screwing the cap 28 to the inlet end part of the column body 21. As a result, the adsorbent 3 is filled into the adsorbent filling space 20 of the column body 21, and an adsorption device 1 can be obtained in which the liquid-tightness of the adsorbent filling space 20 is ensured by the column body 21 and the lid parts 22, 23. By going through the above-mentioned steps [S1B] to [S5B], the adsorption device 1 is manufactured.

[0119] In the present embodiment, the column combination 250 in which the extension column 200 is connected to the column 2 is described as a case in which the adsorbent 3 is filled into the adsorbent filling space 20 of the column body 21. However, for example, when a filling liquid 66 containing the adsorbent 3 is supplied to the adsorbent filling space 20 of the column body 21 using the pump 60, the connection of the extension column 200 to the column 2 can be omitted.

[0120] In this case, the steps [S2B] and [S3B] can be omitted, and in the step [S4B], the column 2 having the lid portions 22 and 23 attached to the column body 21 is oriented so that the lid portion 22 is facing vertically downward, and the filling liquid 66 containing the adsorbent 3 is supplied to the adsorbent filling space 20 of the column body 21 via the inlet pipe 24 of the lid portion 22.

[0121] Furthermore, in the present embodiment, in column 2, the lid portion 22 side is the inflow side through which liquid flows into the adsorbent filling space 20, and the lid portion 23 side is the outflow side through which liquid flows out of the adsorbent filling space 20; however, this relationship can also be reversed, so that liquid flows in from the lid portion 23 side and flows out from the lid portion 22 side via the adsorbent filling space 20.

[0122] In another preferred embodiment of the present invention, instead of filling the adsorbent filling space 20 with the adsorbent 3 as in the above steps [S1B] to [S5B], the adsorbent filling space 20 may be filled in a weightless environment. Since heavy particles do not settle in a weightless environment, it is possible to appropriately suppress or prevent the variation in particle size distribution of the powder in the column without necessarily performing the above steps [S1B] to [S5B]. Alternatively, when filling the adsorbent filling space 20 with the adsorbent 3, a filling liquid with a very high viscosity can be used to prevent or suppress the settling of heavy particles.

[0123] Although the suction device according to the embodiment of the present invention and the method for manufacturing the suction device according to the embodiment of the present invention have been described above, the present invention is not limited to this.

[0124] For example, in the adsorption device according to the embodiment of the present invention, each component can be replaced with any component that can exert a similar function, or any component can be added. Also, in the manufacturing method of the adsorption device according to the embodiment of the present invention, one or more steps can be added for any purpose.

[0125] In the above embodiment, the column body of the column and the column passing section of the extension column are each described as being cylindrical, but the column body and the column passing section may be cylindrical in shape, and may be polygonal, such as a hexagon or heptagon. EXAMPLES

[0126] Next, specific examples of the present invention will be described. However, the present invention is not limited to the descriptions in these examples.

[0127] 1. Examination of the packing property of the adsorbent in the column (Reference example 1) [R1-1] First, a column 2 having a column body 21 with an inner diameter of φ50 mm and a column length of 230 mm and an extension column 200 having a column passing section 221 with an inner diameter of φ50 mm and a column length of 230 mm were prepared. The connecting section 229 of the extension column 200 was attached to the inlet end of the column body 21 of the column 2 by screwing, thereby obtaining a column connector 250.

[0128] [R1-2] As the adsorbent 3, 248 g of sintered powder of hydroxyapatite (manufactured by BIO-RAD, CHT 40 μm Type I) was weighed out, and this adsorbent 3 was dispersed in 737 mL of 0.2 mM sodium phosphate buffer by stirring at 25° C. to prepare a slurry 30.

[0129] [R1-3] As shown in Fig. 4, the column coupler 250 was arranged so that the column 2 was vertically downward and the extension column 200 was vertically upward. Thereafter, the slurry 30 was supplied from an opening portion that opened at the upper end portion of the column passing portion 221 provided in the extension column 200 to the adsorbent passing space 220 provided in the column passing portion 221, thereby filling the adsorbent passing space 220 with the slurry 30. Thereafter, a cap 228 was attached to the upper end portion of the column passing portion 221 by screwing.

[0130] [R1-4] Next, the column combination body 250 was inverted and stirred vertically multiple times to suspend the adsorbent 3 that had settled due to gravity, and the column combination body 250 was maintained in an orientation in which the column 2 was vertically upward and the extension column 200 was vertically downward. At the same time, the supply of water as the filling liquid 66 contained in the container 65 was started using the pump 60 to the adsorbent passing space 220 of the column passing section 221 via the inlet pipe 224 of the cap 228.

[0131] At this time, the flow rate of water through the adsorbent passing space 220 of the column passing portion 221 using the pump 60 was set to 160 mL / min.

[0132] Then, water was continuously supplied to the adsorbent passing space 220 at a flow rate of 160 mL / min for 30 minutes, and the thickness of the adsorbent 3 filled in the adsorbent passing space 220 from the inlet end to the outlet end of the column body 21 was measured.

[0133] (Reference examples 2 and 3) In the step [R1-4], the flow rate of water and the water supply time when water was supplied to the adsorbent passing space 220 of the column passing section 221 using the pump 60 were changed as shown in Table 1, respectively. In the same manner as in Reference Example 1, the thickness of the adsorbent 3 filled in the adsorbent passing space 220 from the inlet end to the outlet end of the column body 21 was measured.

[0134] Table 1 shows the thicknesses of the adsorbent 3 filled in the adsorbent passing space 220 measured in Reference Examples 1 to 3.

[0135] [Table 1]

[0136] As is clear from Table 1, for a column body 21 having an inner diameter of φ50 mm, by setting the flow rate to 80 mL / min or more, the adsorbent 3 can be filled into an extension column 200 having an adsorbent passing space 220 with a column length of approximately 180 mm or less, and it has become clear that the water flow rate can be appropriately set depending on the column inner diameter, column length, filling time, etc. (for example, if the column length is approximately 200 mm, the water flow rate can be set to 160 mL / min).

[0137] 2. Study of particle size distribution of adsorbent in each region of the adsorption device 2-1. Manufacturing of adsorption equipment Example 1 [E1-1] First, a column combination 250 was obtained in the same manner as in the above step [R1-1].

[0138] [E1-2] Next, a slurry 30 was prepared in the same manner as in the above-mentioned step [R1-2]. [E1-3] Next, in the same manner as in the above-mentioned step [R1-3], the adsorbent passing space 220 was filled with the slurry 30, and then the cap 228 was attached to the upper end of the column passing portion 221.

[0139] [E1-4] Next, in the same manner as in the above-mentioned step [R1-4], the column combination body 250 was inverted and stirred several times in the vertical direction to suspend the adsorbent 3 that had settled due to gravity, and the column combination body 250 was maintained in an orientation in which the column 2 was vertically upward and the extension column 200 was vertically downward. At the same time, the supply of water as the filling liquid 66 contained in the container 65 was started using the pump 60 to the adsorbent passing space 220 of the column passing section 221 via the inlet pipe 224 of the cap 228.

[0140] At this time, the flow rate of water through the adsorbent passing space 220 of the column passing portion 221 using the pump 60 was set to 160 mL / min.

[0141] [E1-5] Next, the supply of water to the adsorbent passing space 220 of the column passing section 221 using the pump 60 was continued until the adsorbent 3 rose sequentially through the adsorbent passing space 220 of the column passing section 221 and the adsorbent filling space 20 of the column body 21, until the adsorbent filling space 20 of the column body 21 was filled with almost the entire amount of the adsorbent 3. Thereafter, the column was returned to its original state and the entire amount was filled.

[0142] [E1-6] Next, the operation of the pump 60 was stopped to terminate the supply of water to the adsorbent passing space 220 of the column passing section 221, and then the connection part 229, which was attached by screwing to the inlet end of the column body 21 of the column 2, was detached from the column body 21, thereby detaching the extension column 200 from the column 2. Then, the lid part 22 was attached to the inlet end of the column body 21 from which the connection part 229 had been detached, by screwing the cap 28 to the inlet end of the column body 21, thereby obtaining the adsorption device 1 of Example 1.

[0143] Comparative Example 1 The adsorption apparatus 1 of Comparative Example 1 was obtained in the same manner as in Example 1, except that in the steps [E1-4] and [E1-5], the inversion of the column connection body 250 so that the column 2 is vertically upward and the extension column 200 is vertically downward was omitted, and water was supplied to the adsorbent passing space 220 of the column passing section 221 using the pump 60.

[0144] 2-2. Measurement of particle size distribution of adsorbent in each region of the column of the adsorption device In the adsorption apparatus of Example 1 and the adsorption apparatus of Comparative Example 1, after the cover portion 22 was detached from the column body 21, the column body 21 was maintained horizontally and water was supplied from the cover portion 23 by the pump 60, whereby the adsorbent 3 filled in the adsorbent filling space 20 was removed from the column 2.

[0145] Thereafter, about 1 g of this adsorbent 3 was collected using a spatula from the center of the end (one end) which is the first region a1 on the inflow pipe 24 side of the column body 21, about 1 g of the center of the central part (midpoint) which is the second region a2, and about 1 g of the center of the end (other end) which is the third region a3 on the outflow pipe 25 side of the column body 21, and each was collected in a container containing water.

[0146] Then, the particle size distribution of the adsorbent 3 present at the positions corresponding to each region (first region a1, second region a2, and third region a3) was measured using an MT3300EX2 (manufactured by Microtrack Bell) under the following conditions: number of measurements: 2, measurement time: 30 seconds, distribution display: volume, particle refractive index: 1.65, solvent: water, solvent refractive index: 1.333.

[0147] In addition, the particle size distribution of the adsorbent 3 present at the positions corresponding to each region (first region a1, second region a2, and third region a3) in the adsorption devices of Example 1 and Comparative Example 1 was measured twice, and the particle size distribution of the adsorbent 3 was calculated by finding the average value from the measurement values ​​for the two lots. The results are shown in Table 2.

[0148] FIG. 6 is a graph showing the particle size distribution of the adsorbent 3 present at the position corresponding to the first region a1 in the adsorption device of Example 1.

[0149] [Table 2]

[0150] As is clear from Table 2, in the adsorption apparatus of Example 1, in the steps [E1-4] and [E1-5], the column connection body 250 was in a position in which the column 2 was vertically upward and the extension column 200 was vertically downward, and water was supplied to the adsorbent passing space 220 of the column passing section 221 using the pump 60 to fill the adsorbent 3 in the adsorbent filling space 20. As a result, when the most frequent particle size in the particle size distribution of the adsorbent 3 present in the first region a1 is A1 [μm] and the most frequent particle size in the particle size distribution of the adsorbent 3 present in the third region a3 is A3 [μm], the relationship 0.72≦A1 / A3≦1.38 was satisfied. That is, an adsorption apparatus in which the variation in the particle size distribution of the adsorbent 3 in the column 2 was appropriately suppressed was obtained in Example 1.

[0151] In contrast to this, in the adsorption apparatus of Comparative Example 1, in the steps [E1-4] and [E1-5], the inversion of the column combination body 250 in which the column 2 is vertically upward and the extension column 200 is vertically downward is omitted, and water is supplied to the adsorbent passing space 220 of the column passing section 221 using the pump 60 to fill the adsorbent filling space 20 with the adsorbent 3, resulting in the result that the relationship 0.72≦A1 / A3≦1.38 cannot be satisfied. That is, the adsorption apparatus obtained in Comparative Example 1 showed the result that the variation in particle size distribution of the adsorbent 3 in the column 2 cannot be suppressed.

[0152] In addition, when bovine serum albumin was purified and isolated using the adsorption devices of Example 1 and Comparative Example 1, respectively, the adsorption device of Example 1 was found to be capable of repeatedly performing purification and isolation multiple times with superior accuracy compared to the adsorption device of Comparative Example 1, i.e., the adsorption device of Example 1 had improved separation performance compared to the adsorption device of Comparative Example 1. [Explanation of symbols]

[0153] 1 Adsorption device 2 Columns 3. Adsorbents 4 Filter material 5 Filter material 20 Adsorbent filling space 21 Column body 22 Lid 23 Lid 24 Inflow pipe 25 Outflow pipe 26 Lid 27 Lid 28 Cap 29 Cap 30 Slurry 41 Flow Path 51 Flow Path 60 Pump 65 Container 66 Filling liquid 200 Extension Column 220 Adsorbent passage space 221 Column passage 224 Inflow pipe 228 Cap 229 Connecting part 250 Column Connection a1 1st area a2 2nd area a3 3rd area

Claims

1. A column body having a cylindrical shape and an adsorbent filling space therein; a first port provided at one end of the column body and having a first flow path through which a liquid flows; a second port provided at the other end of the column body and having a second flow path through which a liquid flows; an adsorbent filled in the adsorbent filling space, The adsorbent includes a powder consisting of an aggregate of fine particles having a non-uniform particle size, An adsorption device characterized in that, when the most frequent particle size in the particle size distribution of the powder present in a first region that is one end of the adsorbent filling space is A1 [μm], and the most frequent particle size in the particle size distribution of the powder present in a third region that is the other end of the adsorbent filling space is A3 [μm], the relationship of 0.72≦A1 / A3≦1.38 is satisfied.

2. 2. The adsorption device according to claim 1, wherein, with the first lid portion positioned vertically above the second lid portion, a sample liquid containing an adsorbent is supplied to the adsorbent filling space via the first flow path of the first lid portion, and the adsorption of the adsorbent is performed by the powder.

3. 3. The adsorption device according to claim 2, wherein when a mode particle size in a particle size distribution of the powder in a second region located at a midpoint between the one end and the other end is A2 [μm], 0.72≦A1 / A2≦1.38 is satisfied.

4. 4. The adsorption device according to claim 3, wherein when the frequency of the most frequent particle size in the particle size distribution of the powder in the second region is B2 (%), 0.80≦B1 / B2≦1.15 is satisfied.

5. A first step of preparing a column including a cylindrical column body having an adsorbent-filled space therein, a first port provided at one end of the column body and having a first flow path through which a liquid flows, and a second port provided at the other end of the column body and having a second flow path through which a liquid flows; a second step of filling the adsorbent filling space of the column with a composition including an adsorbent containing a powder consisting of an aggregate of fine particles having a non-uniform particle size and a first liquid; and a third step of supplying a second liquid to the adsorbent filling space via the first flow path of the first port while the first port side of the column is positioned vertically downward.

6. Prior to the second step, preparing an extension column including: a column passing portion having a cylindrical shape and an adsorbent passing space therein; a cover body provided at one end of the column passing portion and having a third flow path through which a liquid flows; and a connecting portion provided at the other end of the column passing portion and connectable to the one end side of the column main body; 6. The method for manufacturing an adsorption device according to claim 5, further comprising the step of connecting the other end of the column passing portion to the one end of the column body from which the first port is detached, via the connecting portion.

7. The adsorption device obtained by the third step is:

7. The method for manufacturing an adsorption device according to claim 5 or 6, wherein, when the most frequent particle size in the particle size distribution of the powder present in one end region of the adsorbent filling space is A1 [μm] and the most frequent particle size in the particle size distribution of the powder present in the other end region of the adsorbent filling space is A3 [μm], 0.72≦A1 / A3≦1.38 is satisfied.

Images